Cyano substituted malanilate biocides

ABSTRACT

itut Neorganicheskoi Khimii Riga SU 03

United States Patent i 1 [111 3,860,627

Large Jan. 14, 1975 CYANO SUBSTITUTED MALANILATE 3,254,108 5/1966Maggiulli et al 260/465 BIOCIDES Primary ExaminerLewis Gotts AssistantExaminerD0lph H. Torrence Attorney, Agent, or FirmDaniel C. Block [57]ABSTRACT This application relates to a biocide active compound havingthe following formula II ll c n o-c-c zn-c-nn c1 GEN 1 Claim, N0Drawings 1 CYANO SUBSTITUTED MALANILATE BIOCIDES DESCRIPTION OF THEINVENTION This invention is directed to a compound which may begenerally described as a cyano substituted malanilate compound that isactive as a biocide. The compound of the present invention has thefollowing formula 3 l? c a o-c-ca-c-mi 01 The above compound can beprepared by combining 11.3 g. (0.10M) ethyl cyano acetate and 18.8 g.(0.10M) of 3,4-dichlorophenyl isocyanate in a reaction vessel. Whilethese components were stirred, small portions of triethylamine was addedslowly. An exothermic reaction took place which required maintaining thetemperature at a range of 30-40C. during addition. After the addition of10.1 g. (0.10M) of triethylamine, the mixture was heated to 80C. withstirring for 30 minutes. Thereafter, the mixture was cooled anddissolved in acetone The resulting solution was mixed into a rapidlystirred dilute hydrochloric acid solution. The precipitate is filteredand washed with water, dried in an oven overnight to yield 26.0 g. of awhite solid.

BIOCIDE TESTING PROCEDURES Tubes of sterilized nutrient and malt extractbroth are prepared. Aliquots of the toxicant, dissolved in anappropriate solvent, are injected through the stopper, into the broth,to provide concentrations ranging from 50 ppm downward. The testorganisms consist of two fungi, Aspergillus niger (A.n.) van Tieghem andPenicillium italicum (P.i.) Wehmer, and two bacteria, Escherichia coli(E.c.) Migula and Staphylococcus aureus (S.a.) Rosenbach. Three drops ofa spore suspension of each of the fungi are injected into the tubes ofmalt broth and three drops of the bacteria are injected into thenutrient broth. One week later, the growth of each organism is observedand effectiveness of the chemical is recorded as the lowestconcentration in ppm which provides 50 percent inhibition of growth ascompared to untreated inoculated tubes. The results of these tests aretabulated in Table 1.

IN VITRO AGAR SCREENING TESTS This test measures the bactericidal,fungicidal and algaecidal properties of a compound when in contact withgrowing bacteria, fungi or algae in an artificial medium. The test isconducted by adding 20 ml. portions of a suitable warm sterile agarsolution into 20 X 100 mm. Petri dishes. Then, the test compound, in 0.5percent acetone solution, is added to the Petri dishes at levels of 1, land 50 ag/ml. and mixed with the warm mobile agar solution. The treateda'gar mixture is then allowed to come to room temperature and solidify.Cells of the chosen organism are streaked on the surface of thesolidified agar and are then incubated for such lengths of time thatuntreated samples containing no toxicant show luxurious growth typicalof the particular organism. This time varies from 24 'hours to one week,depending on the particular organism. The fungi are incubated at 30C.and the bacteria are incubated at 37C. The algae are incubated at roomtemperature under artificial light. Nutrient agar is used as the mediumin this test for the bacteria. Potato dextrose agar is used as themedium for the fungi with the exception of Pullularia pullulans andTrichophyton mentagmphytes for which Emmons agar is used. A modifiedJack Meyers agar is used for the growth of the algae.

The extent of growth is noted at the end of the incubation period.

Representative organisms used in this test are as follows: BacteriaEnterobacter aerogertes Bacillus cereus Pseudomonas aeruginosaBrevibacterium ammoniagenes Staphylococcus aureus Escherichia coli FungiAsperigillus flavus Aspergillus fumigatas Aspergillus niger Aspergilluso'ryzae Penicillium italicum Penicillium expansum Penicillium sp.

Rhizopus stolonifer Trichophyton menlagrophytes Pullularia pullulansAlgae Scenedesmus obliquus Chlorella pyrerzoidosa TABLE II In Vitro AgarScreening Tests Minimum Inhibitory Concentration. ug/ml.

( indicates partial control at this concentration not tested greaterthan suLFA'ra EouciNo BACTERIA IN VITRO TEST This test measures thebactericidal properties of a compound when in contact with a sulfatereducing bacteria, specifically Desulfovibrio desulfuricans. The test isconducted by dissolving the test compound in acetone to give an 0.5percent solution. This toxicant is added to vials containing sterileSulfate API broth with tryptone under anaerobic conditions at suchlevels to give final toxicant concentrations of l, 5, and 50 pg/ml. ofsolution. An inoculant solution of 0.5 ml. of the growing organism,Desulfovibrio desulfuricans, is added to the vials followed bysufficient sterile distilled water to give a total of 10 ml. solution inthe vials. The vials are incubated at room temperature for 3 to 5 daysuntil untreated controls show growth of the organism as indicated by theblack color development in the vials.

The following is a summary of the minimum inhibitory concentrationnecessary to control the organism.

TABLE III Compound Desulfovibrio desulfuricans 5 The compound of thisinvention is generally embodied into a form suitable for convenientapplication. For example, the compound can be embodied into pesticidalcompositions which are provided in the form of emulsions, suspensions,solutions, dusts and aerosol sprays. In general, such compositions willcontain, in addition to the active compound, the adjuvants which arefound normally in pesticide preparations. In these compositions, theactive compound of this invention can be employed as the sole pesticidecomponent or it can be used in admixture with other compounds havingsimilar utility. The pesticide compositions of this invention cancontain, as adjuvants, organic solvents, such as sesame oil, xylenerange solvents, heavy petroleum, etc.; water; emulsifying agents;surface active agents; talc; pyrophyllite; diatomite; gypsum; clays;propellants, such as dichlorodifluoromethane, etc. If desired, however,the active compound can be applied directly to feedstuffs, seeds, etc.upon which the posts feed. When applied in such a manner, it will beadvantageous to use a compound which is not volatile. In connection withthe activity of the presently disclosed pesticidal compounds, it shouldbe fully understood that it is not necessary that they be active assuch. The purposes of this invention will be fully served if thecompound is rendered active by external influences, such as light or bysome physiological action which occurs when the compound is ingestedinto the body of the pest.

The precise manner in which the pesticidal compositions of thisinvention are used in any particular instance will be readily apparentto a person skilled in the art. Generally, the active pesticide compoundwill be embodied in the form of a liquid composition; for example, anemulsion, suspension, or aerosol spray. While the concentration of theactive pesticide in the present compositions can vary within rather widelimits, ordinarily the pesticide compound will comprise not more thanabout 15.0 percent by weight of the composition. Preferably, however,the pesticide compositions of this invention will be in the form ofsolutions or suspensions containing about 0.1 to 1.0 percent by weightof the active pesticide compound.

What is claimed:

1. A compound having the following formula:

II II. C1 C H O C C IH C NH

